Description
High-yield Taq PCR master mix with loading dye
GenieTaq High Yield 2X Master Mix (Blue) is the dye-containing format of the high-yield GenieTaq mix. It delivers high yield and amplifies up to 10 kb from genomic DNA and up to 15 kb from plasmid DNA and λDNA. A blue tracking dye lets you load products directly onto a gel.
The mix contains GenieTaq High Yield DNA Polymerase, dNTPs and an optimised buffer system; just add primers and template. Compared with standard Taq DNA polymerase it offers higher fidelity, stronger amplification and higher yield, and the premixed format reduces pipetting for better throughput and reproducibility. Protective agents keep activity stable through repeated freeze-thaw cycles. Products carry a 3'-dA overhang for direct T-vector cloning and are also compatible with seamless and TOPO cloning kits. The mix is designed for conventional PCR. For research use only.
Key Features
| High performance | Fidelity ~6× higher than Taq DNA polymerase, with higher amplification yield. | |
| Long targets | Amplifies up to 10 kb genomic DNA and up to 15 kb plasmid DNA and λDNA. | |
| Convenient operation | Ready-to-use mix with tracking dye for direct gel loading of PCR products. | |
| Good stability | Stable performance after 50 freeze-thaw cycles. | |
| Cloning-ready products | 3'-dA overhangs allow direct TA cloning into T vectors. |
Specifications
| Format | 2X ready-to-use master mix with tracking dye (polymerase, dNTPs, optimised buffer) |
| Application | Conventional PCR |
| Fidelity | ~6× higher than Taq DNA polymerase |
| Maximum fragment length | 10 kb (genomic DNA); 15 kb (plasmid DNA and λDNA) |
| Product ends | 3'-dA overhang |
| Stability | Stable after 50 freeze-thaw cycles |
| Shelf life | 2 years |
Components
| Components | Amount |
| GenieTaq High Yield 2X Master Mix (Blue) | 5 × 1 ml / 15 × 1 ml / 50 × 1 ml |
Storage
Store at -30 ~ -15°C.
- Prepare the reaction system
- Set up a 50 μl reaction as follows:
| Components | Volume |
| ddH2O | To 50 μl |
| GenieTaq High Yield 2X Master Mix (Blue) | 25 μl |
| Primer 1 (10 μM) | 2 μl |
| Primer 2 (10 μM) | 2 μl |
| Template DNA* | x μl |
*Recommended template amount in a 50 μl reaction:
| Template type | Amount |
| Animal and plant genomic DNA | 0.1–1 μg |
| E. coli genomic DNA | 10–100 ng |
| cDNA | 1–5 μl (≤1/10 of the total PCR volume) |
| Plasmid DNA | 0.1–10 ng |
| λDNA | 0.5–10 ng |
- Run the reaction program
| Step | Temperature | Time | Cycles |
| Initial denaturationa | 95°C | 3 min | 1 |
| Denaturation | 95°C | 15 sec | 30–35 |
| Annealingb | 60°C | 20 sec | 30–35 |
| Extension | 72°C | 60 sec/kb | 30–35 |
| Final extension | 72°C | 5–10 min | 1 |
a These initial denaturation conditions suit most reactions. For templates with complex structure, extend the initial denaturation to 5–10 min.
b Adjust the annealing temperature to the primer Tm, generally 3–5°C below Tm. For complex templates, adjust the annealing temperature and extend the extension time for efficient amplification.
- Analyse the products
- The mix contains loading dye, so load products directly onto an agarose gel after the reaction.
Q1: What should I do if there is no amplification or the yield is low?
- Optimise primer design; run an annealing temperature gradient to find the optimum; increase primer concentration; extend the extension time; increase cycles to 35–40; use high-purity template; and for crude samples decrease the input, while for other samples increase the amount within the recommended range.
Q2: How can I reduce nonspecific products or smeared bands?
- Optimise primer design; raise the annealing temperature up to 65°C in 2°C steps; decrease the final primer concentration to 0.2 μM; shorten the extension time if nonspecific bands are larger than the target; reduce cycles to 25–30; use high-purity template; and adjust the template amount to the recommended range.
Q3: Can I load the PCR product straight onto a gel?
- Yes. The mix contains loading buffer with a tracking dye, so products can be loaded directly for electrophoresis after the reaction.
Q4: Are the products suitable for TA cloning?
- Yes. Products carry a 3'-terminal adenine and can be cloned into T vectors. They are also compatible with seamless and TOPO cloning kits.
